Staphylococcal-derived superantigen enhances peanut induced Th2 responses in the skin.

BACKGROUND: The allergen-induced activation and expansion of IL-4 producing T helper type 2 (Th2) cells is a key event in the initiation and progression of allergic disease. Intriguingly, concomitant early childhood staphylococcal skin infections are being increasingly implicated in the allergen-induced switch of primary T cell responses towards the Th2 phenotype. OBJECTIVE: We sought to identify whether or not staphylococcal-derived superantigen can influence the primary T cell response in the skin to food allergens with a view to determining whether such exposures create the immune pathology that predisposes to the development of food allergy. METHODS: Using a novel Th2 reporter model, we determined the ability of the staphylococcal superantigen (SEB) to influence priming in the skin of IL-4 expressing Th2 cells by peanut extract (PE). Factors including the effect of SEB on the magnitude of the Th2 response in the skin draining lymph nodes, T cell receptor V region usage and the influence of endotoxin were evaluated. RESULTS: Primary exposure to PE and SEB lead to significantly enhanced PE specific Th2 responses when the mice were subsequently exposed to PE alone. The enhancement of the Th2 response was dependent on the Vß-binding properties of the SEB, but was not affected by endotoxin-mediated TLR-4 effects or strain differences in the mice. CONCLUSIONS AND CLINICAL RELEVANCE: These results identify that in the skin environment, the presence of SEB can significantly increase the numbers of allergen-induced Th2 cells which develop in response to subsequent allergen exposure. These data highlight the process by which individuals may become pathologically sensitized to food allergens in early life.

Protective immunity to nematode infection is induced by CTLA-4 blockade.

The recent observation that neutralization or genetic deletion of the T lymphocyte receptor CTLA-4 allows enhanced T cell reactivity offers new opportunities for immunotherapy against infectious agents. We used a neutralizing antibody to block CTLA-4 interaction with its ligands CD80 and CD86 during infection of mice with the nematode, Nippostrongylus brasiliensis. CTLA-4 blockade greatly enhanced and accelerated the T cell immune response to N. brasiliensis, resulting in a profound reduction in adult worm numbers and early termination of parasite egg production. The ability of CTLA-4 blockade to accelerate primary immune responses to a protective level during an acute infection indicates its potential as an immunotherapeutic tool for dealing with infectious agents.

Th2-dependent airway eosinophilia is regulated by preproenkephalin.

We have used preproenkephalin (PPNK)-deficient mice to study the role of PPNK in the development of airway inflammation. Airway eosinophilia was established by either sensitization followed by airway challenge with OVA or by infection with Nippostrongylus brasiliensis. Both models induce a strong Th2 immune response, characterized by an IL-5-dependent airway eosinophilia. We observed that although the accumulation of lymphocytes in the airways of PPNK-deficient mice was similar to that induced in control mice, IL-5 production and eosinophil infiltration were reduced. We conclude from this work that PPNK has a role in enhancing Th2 cell function and that this molecule may be an important target in asthma immunotherapy.

Application of enzymeimmunoassay to measure oestrone sulphate concentrations in cow's milk during pregnancy.

The characteristics of antigen- and antibody-coated enzymeimmunoassay (EIA) formats to measure oestrone sulphate (OS) were studied using a murine monoclonal antibody as the primary antibody. In an antigen-coated format the most sensitive EIA (9 fmol/well) was achieved using 6-ketoestrone-6-O-carboxymethyloxime (OCMO) coupled to bovine serum albumin (BSA), as the coating antigen, and horseradish peroxidase (HRP), as the enzyme label. In an antibody-coated format, comparable sensitivity could be achieved using HRP conjugated to either OCMO, oestrone-3-glucuronide (OG) or oestrone-3-hemisuccinate (OHS) as the steroid 'tracer'. In both the antigen- and antibody-coated formats replacing HRP with alkaline phosphatase (AP) markedly aggravated the assay sensitivity. The antigen-coated EIA format was used to measure OS concentrations in cow's milk directly without an initial defatting step, and a progressive increase in OS concentrations in milk as pregnancy progressed was observed. Median OS concentrations rose from 1.1 nmol/l at days 70-99 of pregnancy (n = 44) to 3.2 nmol/l at days 140-160 (n = 92). Oestrone sulphate concentrations in milk from non-pregnant cows (n = 51) ranged from non-detectable to 1.3 nmol/l with a median value of 0.4 nmol/l. Only 5% of cows 120 or more days pregnant had milk OS concentrations within the range of values found in milk from non-pregnant cows. Accurate discrimination of non-pregnant and pregnant cows can thus be achieved on the basis of OS concentrations in milk samples taken at least 120 days after mating/insemination. This EIA for OS may have a role in the dairy industry as an alternative non-invasive means of determining pregnancy status in cows.

Evaluation of antibody- and antigen-coated enzymeimmunoassays for measuring oestrone-3-glucuronide concentrations in urine.

A monoclonal antibody to oestrone-3-glucuronide (OG) was generated and incorporated into antigen- and antibody-coated competitive enzymeimmunoassays (EIAs) using OG-, 6-ketoestrone-6-O-carboxymethyl-oxime (OCMO) and oestrone-3-hemisuccinate (OHS) as the steroid coating antigens or 'tracers' in each format respectively. In the coated-antigen format, standard curves with the lowest mean values (pg/well) for sensitivity (1.1 +/- 0.1), mid-point (ED50; 8.2 +/- 0.7) and high-point (ED20; 31 +/- 2) were obtained using OCMO coupled to bovine serum albumin (BSA) as the coating antigen, and horseradish peroxidase (HRP) as the enzyme label coupled directly to the monoclonal antibody. Standard curves generated using enzyme-labelled OG, OCMO and OHS as 'tracers' in the antibody-coated EIA format were all similar, but had higher mean sensitivity, ED50 and ED20 values than those obtained in the optimal coated-antigen format. In both EIA formats alkaline phosphatase (AP) was found to be inferior to HRP as an enzyme label. Measurement of OG concentrations in early morning urine samples from women with natural, regular menstrual cycles, using the antigen-coated EIA, demonstrated the characteristic elevation in OG concentrations associated with the onset of the urinary LH surge. This technically straightforward and robust antigen-coated EIA may be of interest to laboratories with a requirement to measure OG concentrations in urine, or other biological samples.

Problems of ethnic groups in our community.

In Australia there are many ethnic groups with varied social and traditional backgrounds. The English migrants, while being predominant in number, are considered to have the least problems, since their emotional and physical environment is similar to that of Australia. Migrants from Western Europe, while having an initial language problem, can be placed in the same category as the English migrant by virtue of Western Europe's comparable industrial and domestic conditions.

Preproenkephalin is a Th2 cytokine but is not required for Th2 differentiation in vitro.

Preproenkephalin (PPNK) mRNA expression has been detected in many cells of the immune system, including monocytes and lymphocytes. In the present paper, the expression of PPNK mRNA in purified CD4+ Th1 and Th2 lymphocyte subpopulations is investigated and correlated with the presence of the opioid neuropeptides leu- and met-enkephalin. We found that PPNK mRNA and met-enkephalin were present at higher levels in the Th2 cultures compared with the Th1 cultures. Lymphocytes from PPNK-deficient mice were then used to look at the role of PPNK in Th2 lymphocyte differentiation. Lymphocytes from these mice could be driven into a Th2 phenotype, suggesting that cultures containing IL-4 do not require PPNK for Th2 differentiation.